柑桔原生质体融合再生叶肉亲本型植株的遗传分析

叶肉亲本（粗柠檬）型植株叶形指数、气孔特征与亲本粗柠檬无异,与同组合体细胞杂种差异显著。染色体计数为二倍体（２ｎ＝２ｘ＝１８）。过氧化物酶（ＰＯＸ）、多酚氧化酶（ＰＰＯ）、谷草转氨酶（ＧＯＴ）同工酶图谱与粗柠檬一致。ＲＡＰＤ分析表明,在具多态性的５４个随机引物中,大多数引物（５２个）上的图谱与粗柠檬相同。但ＯＰＷ－１２上,叶肉亲本型植株含有哈姆林甜检的特征谱带。ＯＰＶ－０４扩增产物显示,叶肉亲本型     

Protoplast-fusion-mediated transfer of organelles from Microcitrus into Citrus and regeneration of novel alloplasmic trees

Summary Iodoacetate-treated Citrus protoplasts from embryogenic nucellar calli of Sour orange (C. aurantium) or from Rough lemon (C. jambhiri) were fused with -irradiated protoplasts from a related genus, Microcitrus. The fused protoplasts were cultured to obtain colonies and micro-calli. Micro-calli derived from these two fusion combinations were isolated, propagated and differentiated into embryos, which subsequently regenerated trees having the morphology of Sour orange or Rough lemon. These intergeneric fusions resulted in mitochondria with novel DNA, indicating recombination between the chondriomes of Citrus and Microcitrus. Chloroplast DNA analyses of fusion-derived embryos indicated that they contained the chloroplasts of either fusion-partner or a mix of these chloroplasts. Later plastome analyses of leaves from fully differentiated plants showed that cybrids having Rough lemon morphology had either Rough lemon or Microcitrus chloroplast DNA, indicating complete sorting out of chloroplasts. Likewise, sorting out of Microcitrus chloroplasts was detected in a cybrid plant having Sour orange morphology.Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250 Israel. No. 2663-E, 1989 series     

哈姆林甜橙与粗柠檬体细胞杂种的育性

对异源四倍体柑桔体细胞杂种“哈姆林甜橙+粗柠檬”及其亲本的花粉活力、花器官发育、花器官形态发生与花粉母细胞减数分裂四分体阶段进行了观测和统计.结果发现“哈姆林甜橙+粗柠檬”的花粉染色活力、萌发率、每花药中花粉粒数均居于其双亲之间,花器官发育及其形态发生具有双亲的特点.但小花粉及花粉母细胞减数分裂过程中形成的不正常四分体比率远远高于其双亲.以体细胞杂种“哈姆林甜橙+粗柠檬”为花粉亲本,与二倍体单胚类型宜昌橙与华农本地早的有性后代杂交,获得了110棵有性后代植株,其中三倍体82棵,二倍体和其它倍性的植株28棵.     

澳洲指橘与柑橘属间原生质体电融合再生二倍体体细胞杂种

电场诱导粗柠檬（CitrusjambhiriLush,2n＝2x＝18）叶肉原生质体与澳洲指橘（MicrocitruspapuanaSwingle,2n＝2x＝18）悬浮系原生质体融合,融合产物培养后再生出丛芽,经试管嫁接得到完整植株。再生植株的细胞学检查表明它们具有18条染色体,为二倍体;植株的叶片形态与叶肉亲本（粗柠檬）一样;用6个10-mer随机引物分析再生植株的杂种特性：在4个引物（OPA－07、OPAN－07、OPE－05和OPA－08）的扩增带型图中,再生植株的带型与粗柠檬完全一样,澳洲指橘的特征带未在植株中出现;在引物OPS-13和引物OPA－04的扩增带型图中,再生植株都具有澳洲指橘的特征带。细胞学和RAPD分析的结果表明,通过对称融合得到了澳洲指橘与粗柠檬的属间二倍体体细胞杂种植株。这是柑橘属间对称融合再生二倍体叶肉亲本类型植株的首例报道。     

Molecular characterization of the nuclear and cytoplasmic genomes of intergeneric diploid plants from cell fusion between Microcitrus papuana and Rough lemon

Simple-sequence-repeat (SSR) and PCR-RFLP were employed to characterize the nuclear and cytoplasmic genomes of intergeneric diploid plants derived from symmetric fusion between Microcitrus papuana Swingle and Rough lemon ( Citrus jambhiri Lush). Three out of five SSR primers distinguished the fusion parents from each other and the regenerated plants showed band profiles completely identical to the leaf parent, Rough lemon. Amplified products from the intergenic regions of cpDNA between trnD -trnT were digested with HaeIII and MspI, and those between trnH -trnK were digested with HinfI, and both the regenerated plants and Rough lemon shared the same band patterns, which were different from the embryogenic parent, M. papuana. With mtDNA, only 2 out of 12 primer pair/restriction enzyme combinations ( nad4 ex 1–2/ TaqI and nad4 ex 1–2/ HindIII) revealed polymorphisms between the fusion parents. With the former combination the regenerated plants showed the same fragment distribution as that of the embryogenic parent, M. papuana, whereas with the latter, a novel band absent in the fusion parents was detected in all of the regenerated plants, suggesting a possible rearrangement. The present research indicates that the plants analyzed were putative cybrids containing nuclear DNA and cpDNA from Rough lemon and mtDNA from M. papuana. Presumed mechanisms leading to the regeneration of diploid hybrid plants following symmetric fusion are discussed herein.     

RAPD-based genetic analysis of offsprings from the sexual cross using allotetraploid citrus somatic hybrid as pollen parent

Thirty-one polymorphic decamer primers were selected to genotype 92 progenies from the cross between Yiben No.4, a monoembryonic diploid F₁ hybrid of Citrus reticulata Blanco cv Huanongbendizao tangerine and C. ichangensis Swingle, and [Hamlin sweet orange + Rough lemon], an allotetraploid somatic hybrid of Citrus sinensis Osbeck cv. Hamlin and C. jambhiri Lush cv. Rough Lemon. χ² (Chi-square) analysis of RAPD markers in the progenies indicated they were randomly transmitted from the four donor parents, without significant difference between the diploids and triploids. However, these progenies were clustered into three major groups using dendrogram constructed by UPGMA, skewed to three parents in certain degrees, 15 (13 triploids and 2 diploids) to Hamlin, 16 (9 and 7) to Yiben No. 4, and 61 (57 and 4) to [Hamlin sweet orange + Rough Lemon] from which genomic contribution was predominant in progenies, respectively.     

Regeneration of Diploid Intergeneric Somatic Hybrid Plants Between Microcitrus a Electrofusion

Leaf-derived protoplasts of Rough lemon ( Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana 5wingle (2n = 2x = 18), with an intention of creating novel germplasm. Six plants were regenerated following protoplasts fusion. Cytological examination demonstrated that they were diploids with 18 chromosomes (2n = 2x = 18). RAPD (random amplified polymorphic DNA) analyses with six arbitrary 10-mer primers showed that the regenerated plants had identical band pattems to those of Rough lemon for primers OPA-07, OPAN-07, OPE-05 and OPA-08, whereas for the other two primers, OPA-04 and OPS-13, bands specific to M. papuana could be detected in the regenerated plants. Cytological and RAPD analysis revealed that the regenerated plants were diploid somatic hybrids between M. papuana and Rough lemon. The putative hybrids were morphologically similar to Rough lemon. This is the first report on production of diploid somatic hybrid plants between citrus with its related genus via symmetric fusion.     

Isozyme and plastid DNA assessment of pedigrees of nineteenth century potato cultivars

Summary Isozyme and ctDNA RFLP patterns were determined for ten historically important potato cultivars (Solanum tuberosum ssp. tuberosuni) in order to relate and confirm their pedigrees. Isozyme polymorphism was detected at 11 of 13 loci examined, whereas only T-type cytoplasm, the predominant ctDNA of S. tuberosum ssp. tuberosum, was observed. Isozyme analysis indicated that potato cultivars previously presumed to be derived from open-pollinated berries of Garnet Chili and Early Rose were in fact the result of hybridizations. In addition, putative pedigrees of Irish Cobbler, White Rose, and Bliss Triumph were not supported. Garnet Chili, the first derivative of Rough Purple Chili, carries allozmyes at Mdh-1 and Pgm-2, which supports the Chilean origin of Rough Purple Chili. The identical ctDNA pattern among the cultivars may indicate a common maternal lineage that traces through Garnet Chili to Rough Purple Chili. The allozyme frequencies estimated from these cultivars provide a base from which subsequent introductions of Solanum species into the ssp. tuberosum gene pool can be assessed.Journal Article No. 000123     

基于AHP与Rough Set的农业节水技术综合评价

由于不同区域自然条件的差异,农业节水技术自身特点及对应用环境条件的要求,导致不同农业节水技术的应用效果存在很大差异,有较大的不确定性,因此,如何筛选合理的评价指标,构建科学、全面的农业节水技术综合评价方法具有十分重要的意义。根据对农业节水技术应用效果的实地调研,应用Delphi法从调查获得的20项评价指标中筛选出了9项农业节水技术综合评价指标。其中节水率、积温和土壤肥力属于生态因子,产投比、劳动力投入和经济投入属于经济因子,可靠性、推广程度和农民认可度为社会因子。基于以上9项指标,构建了农业节水技术综合评价指标体系.综合评价方法(ARM)通过引入经验因子α对应用层次分析法(AHP)和粗糙集(Rough Set)所获得指标权重进行修订,使指标权重更加合理化。同时,分别应用AHP、Rough Set和ARM对甘肃省武威市的地膜覆盖、秸秆覆盖和常规畦田灌溉在大田中的应用效果进行了综合评价。评价结果显示,在对地膜覆盖技术评价中,AHP法过分强调了经济效益的作用(0.44),Rough Set法则强调的是生态效益(0.33)和社会效益(0.32),弱化了经济效益(0.05),ARM修正了以上2种方法的评价结果,获得地膜覆盖的经济效益、生态效益和社会效益,分别为0.36、0.20和0.13。在对秸秆覆盖评价中,与其它两种方法相比,Rough Set法的评价结果存在显著性差异,ARM修正获得的经济效益、生态效益和社会效益,分别为0.09、0.18和0.06。在对常规畦田灌溉评价中,Rough Set法强调了生态效益(0.28),经过修正获得的经济效益、生态效益和社会效益,分别为0.24、0.01和0.13。ARM的评价结果表明,采用地膜覆盖的经济效益最佳;同常规畦田灌溉相比较,地膜覆盖、秸秆覆盖的生态性指数分别比之高0.19和0.17,主要是由于秸秆覆盖具有保墒、增加土壤有机质以及在作物生长后期调节地温的作用,地膜覆盖具有节水、提高苗期土壤温度和促进提前出苗的作用;同秸秆覆盖相比,地膜覆盖和常规畦田灌溉的社会性指数分别高112.12%、18.18%,说明这两种技术在河西半干旱地区具有良好的社会基础。可见,在西北半干旱地区地膜覆盖用于种植玉米的效果最佳,而秸秆覆盖尽管其生态效益较高,但经济效益较低,推广应用存在一定的难度。     

Lateral enzyme topology in the rough endoplasmic reticulum of rat liver

Summary Rough microsomes were subfractionated on the basis of different properties in order to investigate the nature and extent of the enzyme heterogeneity of these vesicles. A discontinuous gradient, containing monovalent cations allowed the separation of a ribosome-poor membrane fraction which was enriched in electron transport enzymes and relatively poor in phosphatases. Zonal centrifugation on a stabilizing gradient separated 3 fractions characterized by enrichment of electron transport enzymes, glucose-6-phosphatase and adenosinetriphosphatase, respectively. An essentially similar pattern was seen when ribosomes were removed with EDTA and the denuded vesicles subfractionated on a sucrose gradient. Rough microsomes from phenobarbitaltreated rats exhibited the same pattern both qualitatively and quantitatively. It appears that electron transport enzymes and two types of phosphatases are heterogeneously distributed among rough microsomal vesicles.This work has been supported by grants from the Swedish Medical Research Council. The authors wish to thank Mrs. Ulla-Britta Torndal for her valuable technical assistance     

RAPD-based genetic analysis of offsprings from the sexual cross using allotetraploid citrus somatic hybrid as pollen parent

Thirty-one polymorphic decamer primers were selected to genotype 92 progenies from the cross between Yiben No.4, a monoembryonic diploid F1 hybrid of Citrus reticulata Blanco cv Huanongbendizao tangerine and C. ichangensis Swingle, and [Hamlin sweet orange + Rough lemon], an allotetraploid somatic hybrid of Citrus sinensis Osbeck cv. Hamlin and C. jambhiri Lush cv. Rough Lemon. χ² (Chi-square) analysis of RAPD markers in the progenies indicated they were randomly transmitted from the four donor parents, without significant difference between the diploids and triploids. However, these progenies were clustered into three major groups using dendrogram constructed by UPGMA, skewed to three parents in certain degrees, 15 (13 triploids and 2 diploids) to Hamlin, 16 (9 and 7) to Yiben No. 4, and 61 (57 and 4) to [Hamlin sweet orange + Rough Lemon] from which genomic contribution was predominant in progenies, respectively     

New method for determining surface roughness of tongue

doi: 10.1111/j.1741‐2358.2011.00509.x New method for determining surface roughness of tongue Objective: The degree of atrophy of the lingual papillae in elderly individuals was evaluated using a quantitative method. Subjects and methods: One hundred and eighty‐two subjects living in nursing homes and 20 healthy adults as controls were studied. To express the degree of atrophy of the lingual papillae quantitatively, lingual surface roughness was determined by taking an impression with silicone dental material. Based on the impressions obtained from the elderly subjects, they were classified by three expert dentists into three groups: Normal, Smooth and Rough. The same determinations were also performed in the 20 healthy controls and compared with 38 of the elderly subjects who had agreement from all of the experts and without the presence of fissures (Normal, n = 6; Smooth, n = 12; Rough, n = 20). Results: The roughness average value for the controls was 65.0 μm, while that for the elderly subjects in the Normal, Smooth and Rough groups was 73.9 μm, 42.2 μm and 94.1 μm, respectively, which were significantly different. Conclusion: The present results indicate that the present technique of obtaining an impression of the tongue surface is simple and reliable for routine evaluation and quantification of the degree of atrophy as well as morphology of the lingual papillae.     

两类柑桔杂种砧木对脐橙(Citrus sinensis Osbeck)幼树生长和叶片糖营养含量年变化的影响

以体细胞杂种红桔+枳和红桔+粗柠檬、有性杂种Troyer枳橙和Swingle枳柚作砧木的耐湿脐橙(CitrussinensisOsbeck)2年生嫁接苗为试材,通过盆栽试验研究了砧木对苗木生长和叶片糖营养含量年变化动态的影响。结果表明:红桔+枳的生长势和花量明显地强于和大于其它砧木,红桔+粗柠檬的生长和花量表现介于2种有性杂种之间。叶绿素a/b和类胡萝卜素含量在不同砧木间无显著差异,而不同砧木显著或极显著地影响叶绿素含量各指标。砧木影响叶片可溶性糖和淀粉含量年变化动态的基本特征。在休眠期,红桔+枳的可溶性糖和淀粉含量都高于2种有性杂种砧木,而红桔+粗柠檬则反之;在生长期,红桔+粗柠檬的可溶性糖和淀粉含量表现出高于有性杂种砧木的趋势,而红桔+枳则介于2种有性杂种砧木之间。同时还重点讨论了2种体细胞杂种砧木的利用价值。     

两类柑桔杂种砧木对脐橙(Citrus sinensis Osbeck)幼树生长和叶片糖营养含量年变化的影响

以体细胞杂种红桔＋枳和红桔＋粗柠檬、有性杂种Troyer枳橙和Swingle枳柚作砧木的耐湿脐橙(Citrus sinensis Osbeck)2年生嫁接苗为试材,通过盆栽试验研究了砧木对苗木生长和叶片糖营养含量年变化动态的影响。结果表明：红桔＋枳的生长势和花量明显地强于和大于其它砧木,红桔＋粗柠檬的生长和花量表现介于2种有性杂种之间。叶绿素a／b和类胡萝卜素含量在不同砧木间无显著差异,而不同砧木显著或极显著地影响叶绿素含量各指标。砧木影响叶片可溶性糖和淀粉含量年变化动态的基本特征。在休眠期,红桔＋枳的可溶性糖和淀粉含量都高于2种有性杂种砧木,而红桔＋粗柠檬则反之;在生长期,红桔＋粗柠檬的可溶性糖和淀粉含量表现出高于有性杂种砧木的趋势,而红桔＋枳则介于2种有性杂种砧木之间。同时还重点讨论了2种体细胞杂种砧木的利用价值。     

The synthesis of α-amylase by rough and in vitro reconstituted rough endoplasmic reticulum derived from rat parotid gland

Summary The isolation of rough and smooth endoplasmic reticulum from rat parotid salivary gland is described. The rough membrane was stripped of its bound ribosomes using the KCl-puromycin method. Rough endoplasmic reticulum was reconstituted from stripped-rough membrane and polyribosomes. The reconstituted rough membrane resembled the native rough membrane in the following aspects: RNA/protein ratio, buoyant density in a continuous sucrose gradient and amino acid incorporation capacity. The in vitro synthesis of -amylase by both rough and in vitro reconstituted rough membrane was demonstrated using SDS polyacrylamide gel electrophoresis. The reconstituted rough membrane could be restripped by KCl-puromycin. The in vitro synthesized -amylase remained associated with the rough or the in vitro reconstituted rough membrane, even after these membranes were stripped of their bound ribosomes.Abbreviations Fp Free polyribosomes - Bp Membrane-bound polyribosomes released by DOC - RM Rough membrane - SM Smooth membrane - RMst Rough membrane stripped - RMrec In vitro reconstituted rough membrane - DOC Sodium deoxycholate     

Introgressive hybridization between Galaxias depressiceps and Galaxias sp D (Teleostei: Galaxiidae) in Otago, New Zealand: Secondary contact mediated by water races

Central Otago (New Zealand) was the focus of an intense period ofgold mining in the late 19th century. Mining relied on high waterpressure for sluicing, often requiring extensive systems of water raceslinking headwaters over large distances. On Rough Ridge, races connectedheadwaters of Otago's two major river systems, the Taieri and theClutha. We employed genetic analysis of freshwater fish(Galaxias) populations to assess the biological implications ofrace-development. The two galaxiid taxa found on Rough Ridge (G. spD and G. depressiceps) are reciprocally monophyletic formtDNA and have three fixed allozyme differences. Rough Ridge aside,G. sp D is apparently absent from the Taieri system and G.depressiceps from the Clutha system. Distributions of mitochondrialDNA, allozyme and microsatellite alleles imply that artificialconnections have facilitated bidirectional introgression: movementof G. sp D alleles into the Taieri system and G.depressiceps alleles into the Clutha. The finding of some identicalcontrol region haplotypes (of both lineages) on either side of thecatchment boundary confirms the recency of genetic exchange. Near theinferred region of contact, samples from Linn Burn (Taieri) are pureG. depressiceps for three diagnostic allozymes and mtDNA.Samples from upper Pool Burn (Clutha) are pure G. sp D forthese same four markers. However, other collections from around thecontact zone represent genetic mixtures forming a cline. We fear thatsimilar human-induced mixing may have happened elsewhere in uplandOtago.     

黄花败酱黄酮类化合物的提取与分析

用70％乙醇提取黄花败酱黄酮类化合物,得到粗黄酮,经聚酰胺纯化,得到精制黄酮。结果表明：黄花败酱干粉中总黄酮含量为2．35％,粗黄酮、精制黄酮中黄酮含量分别为8．81％和21．39％。黄花败黄酮类化合物主要是黄酮和黄酮醇两类,其中含有芦丁成分。     

Discrepancy in compliance between the clinical and genetic diagnosis of choroidal hypoplasia in Danish Rough Collies and Shetland Sheepdogs

Collie eye anomaly (CEA) is a congenital, inherited ocular disorder which is widespread in herding breeds. Clinically, the two major lesions associated with CEA are choroidal hypoplasia (CH) and coloboma, and both lesions are diagnosed based on ophthalmological examination. A 7.8‐kb intronic deletion in the gene encoding non‐homologous end‐joining factor 1 (NHEJ1) has been reported to be the causative mutation underlying CH when present in the homozygous state. In this study, we have investigated the compliance between the clinical and genetic diagnosis of CH in the Danish Rough Collie and Shetland Sheepdog populations. Our results show that the deletion in NHEJ1 is not predictive for CH in the Danish Rough Collie population, whereas the clinical and genetic diagnosis is in accordance with each other in the Shetland Sheepdog population. Based on these results, it can be concluded that the intronic deletion in NHEJ1 is not the causative mutation but, rather, a marker linked to the locus underlying the trait in some populations but linked to both the wild‐type and CH‐causing locus in most dogs in the Danish Rough Collie population.     

Negatively-stained polysomes on rough microsome vesicles viewed by electron microscopy: further evidence regarding the orientation of attached ribosomes

Rough microsomes, derived from rough endoplasmic reticulum of rat liver, were studied by electron microscopy after negative staining, to seek further information about the orientation of ribosomal small and large subunits in bound polysomes. Rough microsomal vesicles were fixed with 2% formaldehyde, centrifuged onto electron-microscopic grid membranes, and were then negatively-stained with 2% phosphotungstic acid. In these preparations, viewed with the electron microscope, flattened rough microsomal vesicles with bound polysomes were sometimes discernible, and the individual ribosomes in the polysomes occasionally showed small and large subunits. The small subunits were uniformly oriented toward the inside of the polysomal curve. The large and small subunits appeared to be alongside one another on the membrane, consistent with the orientation that has been described by Unwin and his co-workers. The boundary between the small and large subunits occurred at approximately the same level in the ribosome where inter-ribosomal strands have been described previously in surface views of bound polysomes in positively-stained electron-microscopic tissue sections. This further confirms the identity of the strands as messenger RNA.This work has appeared in abstract form: Christensen AK (1990)     

GTP-dependent membrane fusion during hepatocarcinogenesis and liver regeneration

Rough microsomes were isolated from homogenates of livers of rats bearing hepatomas as well as from homogenates of livers of rats 24 and 48 h after partial hepatectomy. When incubated in the presence of GTP in a cell-free system to assay membrane fusion these membranes were observed to have a greater capacity (1.4 to 5 fold) for GTP-dependent fusion than homologous membranes from control non-proliferating liver tissue. The enhanced GTP-dependent membrane fusion may reflect changes in membrane properties related to cell proliferation.